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KL Britton HF Rogers Y Asano T Dairi Y Kato TJ Stillman DW Rice 《Acta crystallographica. Section D, Biological crystallography》1998,53(4):124-126
The novel NAD+-linked opine dehydrogenase from a soil isolate Arthrobacter sp. strain 1C belongs to an enzyme superfamily whose members exhibit quite diverse substrate specificites. Crystals of this opine dehydrogenase, obtained in the presence or absence of co-factor and substrates, have been shown to diffract to beyond 1.8 ? resolution. X-ray precession photographs have established that the crystals belong to space group P21212, with cell parameters a = 104.9, b = 80.0, c = 45.5 ? and a single subunit in the asymmetric unit. The elucidation of the three-dimensional structure of this enzyme will provide a structural framework for this novel class of dehydrogenases to enable a comparison to be made with other enzyme families and also as the basis for mutagenesis experiments directed towards the production of natural and synthetic opine-type compounds containing two chiral centres. 相似文献
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A rugged procedure utilizing reversed-phase liquid chromatography with positive-ion electrospray ionization mass spectrometry (LC-MS) along with tandem MS is described for the quantification and confirmation of N-methylpyrrolidinone (NMP) in methanolic extracts of riverine biofilm. The LC-MS method provided a 100-fold improvement in detection limits (2 ng g(-1) with a repeatability of 80-95% based on triplicate analyses) compared to a conventional LC-UV detection procedure and was applicable to quantitative analysis of biofilm samples with little or no clean up. Under low-energy collision induced dissociation (CID) conditions (17 V, laboratory frame of reference, with argon as the collision gas), two product-ions of the [M+H]+ ion were formed at m/z 69 [MH-CH3NH2]+ and m/z 58 [MH-CH3NCH]+ with relative abundances of 30% and 5%, respectively. These CID transitions were used to demonstrate that biofilm uptake of a photocatalytically-generated mixture of NMP was rapid once acclimation was achieved. 相似文献
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A kinetic study of the fluoride ion catalyzed isomerization of a series of 2-aryl1-butenes shows the reactions to be pseudo first order in olefin at constant fluoride ion concentration. The resultant Hammett plot is non-linear witb a concave downward break near =0. A two step mechanism involving formation of a carbanionic intermediate is proposed. A change in the rate limiting step causes the break in the Hammett plot. Carbanion trapping experiments are also reported. 相似文献
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[reaction: see text] The partial reduction of electron-deficient 2,5-disubstituted pyrroles has been developed into a flexible procedure that gives control of relative stereochemistry by variation of the reduction conditions. After the reaction, the pyrroline products were dihydroxylated at C-3,4 to give either the cis or trans isomers; this flexibility means that a variety of polyhydroxylated pyrrolidines can be prepared in a short sequence. Finally, this method was applied to a synthesis of the naturally occurring glycosidase inhibitor DMDP. 相似文献
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Joshua Buse Randy W. Purves Ronald E. Verrall Ildiko Badea Haixia Zhang Christopher C. Mulligan Kerry M. Peru Jonathan Bailey John V. Headley Anas El‐Aneed 《Journal of mass spectrometry : JMS》2014,49(11):1171-1180
The safe use of lipid‐based drug delivery agents requires fast and sensitive qualitative and quantitative assessment of their cellular interactions. Many mass spectrometry (MS) based analytical platforms can achieve such task with varying capabilities. Therefore, four novel high‐throughput MS‐based quantitative methods were evaluated for the analysis of a small organic gene delivery agent: N,N‐bis(dimethylhexadecyl)‐1,3‐propane‐diammonium dibromide (G16‐3). Analysis utilized MS instruments that detect analytes using low‐resolution tandem MS (MS/MS) analysis (i.e. QTRAP or linear ion trap in this work) or high‐resolution MS analysis (i.e. time of flight (ToF) or Orbitrap). Our results indicate that the validated fast chromatography (FC)‐QTRAP‐MS/MS, FC‐ LTQ‐Orbitrap‐MS, desorption electrospray ionization‐collision‐induced dissociation (CID)‐MS/MS and matrix assisted laser desorption ionization‐ToF/ToF‐MS MS methods were superior in the area of method development and sample analysis time to a previously developed liquid chromatography (LC)‐CID‐MS/MS. To our knowledge, this is the first evaluation of the abilities of five MS‐based quantitative methods that target a single pharmaceutical analyte. Our findings indicate that, in comparison to conventional LC‐CID‐MS/MS, the new MS‐based methods resulted in a (1) substantial reduction in the analysis time, (2) reduction in the time required for method development and (3) production of either superior or comparable quantitative data. The four new high‐throughput MS methods, therefore, were faster, more efficient and less expensive than a conventional LC‐CID‐MS/MS for the quantification of the G16‐3 analyte within tissue culture. When applied to cellular lysate, no significant change in the concentration of G16‐3 gemini surfactant within PAM212 cells was observed between 5 and 53 h, suggesting the absence of any metabolism/excretion from PAM212 cells. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
8.
High-power supercontinuum generation in highly nonlinear, dispersion-shifted fibers by use of a continuous-wave Raman fiber laser 总被引:4,自引:0,他引:4
High-power supercontinua are demonstrated in highly nonlinear, dispersion-shifted fibers with a continuous-wave Raman fiber laser. Supercontinuum growth is experimentally studied under different combinations of fiber length and launch power to show output powers as high as 3.2 W and bandwidths greater than 544 nm. Modulation instability (MI) is observed to seed spectral broadening at low launch powers, and the interplay between MI and stimulated Raman scattering plays an important role in the growth of the continuum at high launch powers. The effect on continuum generation of parametric four-wave mixing coupled with the higher-order dispersion properties of the fiber is investigated. 相似文献
9.
N,N-dichloro-o-nitrobenzenesulfonamide (2-NsNCl2) was found to be an effective electrophilic nitrogen source for the direct diamination of alpha,beta-unsaturated ketones without the use of any metal catalysts. The reaction is very convenient to carry out without the protection of inert gases. Molecular sieves (4 A) and temperature were found to play key roles in controlling the formations of 3-trichloromethyl and dichloromethyl imidazoline products (16 examples). The 2-Ns-protection group of the resulting diamine products can be easily cleaved under mild Fukuyama's conditions. A new mechanism hypothesis of [2+3] cyclization and N-chlorination has been proposed to explain the product structures, particularly their regio- and stereochemistry. 相似文献
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